data_5724 #Corrected using PDB structure: 2BRZ_ # #N.B. (Observed* = Observed shift + Offset correction) # #After reference correction, the following residues still #have a CO chemical shift difference (obs*-pred) greater than 5.0ppm: #NUM AA CS Observed* Predicted # 14 K C 178.35 172.56 # #After reference correction, the following residues still #have a N chemical shift difference (obs*-pred) greater than 10.0ppm: #NUM AA CS Observed* Predicted # 14 K N 121.16 111.16 # 39 D N 123.66 110.64 # #The average CS difference between predicted and observed: #HA CA CB CO N HN #N/A N/A N/A -0.45 -0.24 -0.00 # #bmr5724.str.corr chemical shifts have been re-referenced with the following #offsets (these values have been added to the original bmr5724.str file): #HA CA CB CO N HN #N/A N/A N/A -0.45 -0.24 N/A # #The 95% confidence intervals for the above recommended offsets are: # HA CA CB CO N HN # N/A N/A N/A +/-0.30 +/-0.68 +/-0.12 # #The Correlation Coefficients between predicted and observed #chemical shifts are: #HA CA CB CO N HN #N/A N/A N/A 0.572 0.632 0.560 # #The RMSD between predicted and observed* (reference #corrected) chemical shifts are: #HA CA CB CO N HN #N/A N/A N/A 1.033 2.335 0.415 # ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; Backbone 1H, 13C, and 15N Chemical Shift Assignments for R43A Brazzein ; loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Assadi-Porter Fariba M. . 2 Abilgaard Frits . . 3 Blad Heike . . 4 Markley John L. . stop_ _BMRB_accession_number 5724 _BMRB_flat_file_name bmr5724.str _Entry_type new _Submission_date 2003-03-10 _Accession_date 2003-03-10 _Entry_origination author _NMR_STAR_version 2.1.1 _Experimental_method NMR loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 coupling_constants 1 stop_ loop_ _Data_type _Data_type_count '1H chemical shifts' 50 '15N chemical shifts' 50 '13C chemical shifts' 49 'coupling constants' 13 stop_ loop_ _Related_BMRB_accession_number _Relationship 5723 "wt brazzein" 5725 "Ala2ins brazzein" 5726 "D50A brazzein" 5727 "H31A brazzein" 5728 "R33A brazzein" stop_ save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_title ; Correlation of the Sweetness of Variants of the Protein Brazzein with Patterns of Hydrogen Bonds Detected by NMR Spectroscopy ; _Citation_status published _Citation_type journal _MEDLINE_UI_code 22791616 _PubMed_ID 12732626 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Assadi-Porter Fariba M. . 2 Abilgaard Frits . . 3 Blad Heike . . 4 Markley John L. . stop_ _Journal_abbreviation "J. Biol. Chem." _Journal_volume 278 _Journal_issue 33 _Page_first 31331 _Page_last 31339 _Year 2003 loop_ _Keyword "taste" "sweet protein" "hydrogen bond" "J-coupling" "dynamics" "flexibility" "rigidity" "proteinase inhibitor" stop_ save_ ################################## # Molecular system description # ################################## save_system_brazzein _Saveframe_category molecular_system _Mol_system_name Arg43Ala-brazzein _Abbreviation_common brazzein _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label Arg43Ala-brazzein $brazzein stop_ _System_physical_state native _System_oligomer_state monomer _System_paramagnetic no _System_thiol_state 'not reported' save_ ######################## # Monomeric polymers # ######################## save_brazzein _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common brazzein _Name_variant . _Abbreviation_common bz _Mol_thiol_state 'not reported' ############################## # Polymer residue sequence # ############################## _Residue_count 53 _Mol_residue_sequence ; DKCKKVYENYPVSKCQLANQ CNYDCKLDKHARSGECFYDE KANLQCICDYCEY ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 2 ASP 2 3 LYS 3 4 CYS 4 5 LYS 5 6 LYS 6 7 VAL 7 8 TYR 8 9 GLU 9 10 ASN 10 11 TYR 11 12 PRO 12 13 VAL 13 14 SER 14 15 LYS 15 16 CYS 16 17 GLN 17 18 LEU 18 19 ALA 19 20 ASN 20 21 GLN 21 22 CYS 22 23 ASN 23 24 TYR 24 25 ASP 25 26 CYS 26 27 LYS 27 28 LEU 28 29 ASP 29 30 LYS 30 31 HIS 31 32 ALA 32 33 ARG 33 34 SER 34 35 GLY 35 36 GLU 36 37 CYS 37 38 PHE 38 39 TYR 39 40 ASP 40 41 GLU 41 42 LYS 42 43 ALA 43 44 ASN 44 45 LEU 45 46 GLN 46 47 CYS 47 48 ILE 48 49 CYS 49 50 ASP 50 51 TYR 51 52 CYS 52 53 GLU 53 54 TYR stop_ _Sequence_homology_query_date 2004-07-29 _Sequence_homology_query_revised_last_date 2004-07-16 loop_ _Database_name _Database_accession_code _Database_entry_mol_name _Sequence_query_to_submitted_percentage _Sequence_subject_length _Sequence_identity _Sequence_positive _Sequence_homology_expectation_value PDB 1BRZ "Solution Structure Of The Sweet ProteinBrazzein, Nmr, 43 Structures" 98.15 54 98 98 9e-28 PDB 2BRZ "Solution Nmr Structure Of The Sweet ProteinBrazzein, Minimized Average Structure" 98.15 54 98 98 9e-28 PIR S51208 "brazzein - Pentadiplandra brazzeana" 98.15 54 98 98 9e-28 SWISS-PROT P56552 "BRAZ_PENBA Brazzein" 98.15 54 98 98 9e-28 stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $brazzein "Escherichia coli" 562 Eubacteria . Escherichia coli stop_ save_ ######################### # Experimental source # ######################### save_experimental_source _Saveframe_category experimental_source loop_ _Mol_label _Production_method _Host_organism_name_common _Genus _Species _Strain _Vector_name $brazzein 'recombinant technology' . . . . . stop_ save_ ##################################### # Sample contents and methodology # ##################################### ######################## # Sample description # ######################## save_sample_1 _Saveframe_category sample _Sample_type solution loop_ _Mol_label _Concentration_value _Concentration_value_units _Concentration_min_value _Concentration_max_value _Isotopic_labeling $brazzein . mM 2.0 4.0 "[U-100% 13C; U-100% 15N]" 2,2-dimethyl-2-silapentane-5-sulfonate 0.1 mM . . . "sodium azide" 1 mM . . . "succinic acid" 2 mM . . "[U-2H]" stop_ save_ save_sample_2 _Saveframe_category sample _Sample_type solution loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $brazzein 2.0 mM "[U-100% 15N]" stop_ save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_NMR_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model DMX _Field_strength 600 save_ save_NMR_spectrometer_2 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model DMX _Field_strength 750 save_ ############################# # NMR applied experiments # ############################# save_NMR_applied_experiment _Saveframe_category NMR_applied_experiment _Experiment_name ; 15N-HSQC HNCO HNCOCA HNCA 1H-1H-15N TOCSY 1H-1H-15N NOESY long range HNCO ; save_ ####################### # Sample conditions # ####################### save_condition_1 _Saveframe_category sample_conditions loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units pH 5.2 0.2 n/a temperature 310 1 K 'ionic strength' 2 . mM stop_ save_ #################### # NMR parameters # #################### ############################## # Assigned chemical shifts # ############################## ################################ # Chemical shift referencing # ################################ save_chemical_shift_reference _Saveframe_category chemical_shift_reference loop_ _Mol_common_name _Atom_type _Atom_isotope_number _Atom_group _Chem_shift_units _Chem_shift_value _Reference_method _Reference_type _External_reference_sample_geometry _External_reference_location _External_reference_axis _Indirect_shift_ratio DSS H 1 'methyl protons' ppm 0.0 internal direct . . . 1.0 DSS N 15 'methyl protons' ppm 0.0 . indirect . . . 0.101329118 DSS C 13 'methyl protons' ppm 0.0 . indirect . . . 0.251449530 stop_ save_ ################################### # Assigned chemical shift lists # ################################### ################################################################### # Chemical Shift Ambiguity Index Value Definitions # # # # Index Value Definition # # # # 1 Unique # # 2 Ambiguity of geminal atoms or geminal methyl # # proton groups # # 3 Aromatic atoms on opposite sides of the ring # # (e.g. Tyr HE1 and HE2 protons) # # 4 Intraresidue ambiguities (e.g. Lys HG and # # HD protons) # # 5 Interresidue ambiguities (Lys 12 vs. Lys 27) # # 9 Ambiguous, specific ambiguity not defined # # # ################################################################### save_bz_cs_1 _Saveframe_category assigned_chemical_shifts loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $condition_1 _Chem_shift_reference_set_label $chemical_shift_reference _Mol_system_component_name Arg43Ala-brazzein loop_ _Atom_shift_assign_ID _Residue_seq_code _Residue_label _Atom_name _Atom_type _Chem_shift_value _Chem_shift_value_error _Chem_shift_ambiguity_code 1 3 CYS N N 117.46 . 1 2 3 CYS H H 8.45 . 1 3 3 CYS C C 173.95 . 1 4 4 LYS N N 118.76 . 1 5 4 LYS H H 7.94 . 1 6 4 LYS C C 176.35 . 1 7 5 LYS N N 121.06 . 1 8 5 LYS H H 9.01 . 1 9 5 LYS C C 175.65 . 1 10 6 VAL N N 127.76 . 1 11 6 VAL H H 8.79 . 1 12 6 VAL C C 174.75 . 1 13 7 TYR N N 129.86 . 1 14 7 TYR H H 9.23 . 1 15 7 TYR C C 174.75 . 1 16 8 GLU N N 130.66 . 1 17 8 GLU H H 9.02 . 1 20 9 ASN H H 9.26 . 1 21 9 ASN C C 174.05 . 1 22 10 TYR N N 122.36 . 1 23 10 TYR H H 7.51 . 1 24 11 PRO C C 176.55 . 1 25 12 VAL N N 123.76 . 1 26 12 VAL H H 8.07 . 1 27 12 VAL C C 178.15 . 1 28 13 SER N N 117.46 . 1 29 13 SER H H 8.38 . 1 30 13 SER C C 177.45 . 1 31 14 LYS N N 121.16 . 1 32 14 LYS H H 7.89 . 1 33 14 LYS C C 178.35 . 1 34 15 CYS N N 115.56 . 1 35 15 CYS H H 7.86 . 1 36 15 CYS C C 175.35 . 1 37 16 GLN N N 115.36 . 1 38 16 GLN H H 7.37 . 1 39 16 GLN C C 175.05 . 1 40 17 LEU N N 121.36 . 1 41 17 LEU H H 7.32 . 1 42 17 LEU C C 176.05 . 1 43 18 ALA N N 127.66 . 1 44 18 ALA H H 8.54 . 1 45 18 ALA C C 178.55 . 1 46 19 ASN N N 115.96 . 1 47 19 ASN H H 9.07 . 1 48 19 ASN C C 174.75 . 1 49 20 GLN N N 123.36 . 1 50 20 GLN H H 7.68 . 1 51 20 GLN C C 176.15 . 1 52 21 CYS N N 119.06 . 1 53 21 CYS H H 8.45 . 1 54 21 CYS C C 175.55 . 1 55 22 ASN N N 118.66 . 1 56 22 ASN H H 7.97 . 1 57 22 ASN C C 176.85 . 1 58 23 TYR N N 117.96 . 1 59 23 TYR H H 8.11 . 1 60 23 TYR C C 177.75 . 1 61 24 ASP N N 119.76 . 1 62 24 ASP H H 9.07 . 1 63 24 ASP C C 177.55 . 1 64 25 CYS N N 120.06 . 1 65 25 CYS H H 9.30 . 1 66 25 CYS C C 177.35 . 1 67 26 LYS N N 119.86 . 1 68 26 LYS H H 7.68 . 1 69 26 LYS C C 178.55 . 1 70 27 LEU N N 119.86 . 1 71 27 LEU H H 8.27 . 1 72 27 LEU C C 178.13 . 1 73 28 ASP N N 116.76 . 1 74 28 ASP H H 9.01 . 1 75 28 ASP C C 177.95 . 1 76 29 LYS N N 112.06 . 1 77 29 LYS H H 6.63 . 1 78 29 LYS C C 176.45 . 1 79 30 HIS N N 112.96 . 1 80 30 HIS H H 7.03 . 1 81 30 HIS C C 173.65 . 1 82 31 ALA N N 120.46 . 1 83 31 ALA H H 7.66 . 1 84 31 ALA C C 175.15 . 1 85 32 ARG N N 116.66 . 1 86 32 ARG H H 8.30 . 1 87 32 ARG C C 176.35 . 1 88 33 SER N N 109.16 . 1 89 33 SER H H 7.99 . 1 90 33 SER C C 172.85 . 1 91 34 GLY N N 107.46 . 1 92 34 GLY H H 8.61 . 1 93 34 GLY C C 170.55 . 1 94 35 GLU N N 118.62 . 1 95 35 GLU H H 8.28 . 1 96 35 GLU C C 173.95 . 1 97 36 CYS N N 120.36 . 1 98 36 CYS H H 8.81 . 1 99 36 CYS C C 173.45 . 1 100 37 PHE N N 121.06 . 1 101 37 PHE H H 9.04 . 1 102 37 PHE C C 174.55 . 1 103 38 TYR N N 120.86 . 1 104 38 TYR H H 8.60 . 1 105 38 TYR C C 176.15 . 1 106 39 ASP N N 123.66 . 1 107 39 ASP H H 8.57 . 1 108 39 ASP C C 177.55 . 1 109 40 GLU N N 117.56 . 1 110 40 GLU H H 9.01 . 1 111 40 GLU C C 176.85 . 1 112 41 LYS N N 120.46 . 1 113 41 LYS H H 8.23 . 1 114 41 LYS C C 174.95 . 1 115 42 ALA N N 113.06 . 1 116 42 ALA H H 8.30 . 1 117 42 ALA C C 175.55 . 1 118 43 ASN N N 117.45 . 1 119 43 ASN H H 8.62 . 1 120 43 ASN C C 174.45 . 1 121 44 LEU N N 125.76 . 1 122 44 LEU H H 8.45 . 1 123 44 LEU C C 175.25 . 1 124 45 GLN N N 120.96 . 1 125 45 GLN H H 8.47 . 1 126 45 GLN C C 174.25 . 1 127 46 CYS N N 121.76 . 1 128 46 CYS H H 9.20 . 1 129 46 CYS C C 171.65 . 1 130 47 ILE N N 129.16 . 1 131 47 ILE H H 8.73 . 1 132 47 ILE C C 175.35 . 1 133 48 CYS N N 125.96 . 1 134 48 CYS H H 8.49 . 1 135 48 CYS C C 172.45 . 1 136 49 ASP N N 122.36 . 1 137 49 ASP H H 8.80 . 1 138 49 ASP C C 175.95 . 1 139 50 TYR N N 125.16 . 1 140 50 TYR H H 9.60 . 1 141 50 TYR C C 175.35 . 1 142 51 CYS N N 120.46 . 1 143 51 CYS H H 9.50 . 1 146 52 GLU H H 8.30 . 1 149 53 TYR H H 7.74 . 1 stop_ save_ ######################## # Coupling constants # ######################## save_coupling_constants _Saveframe_category coupling_constants loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $condition_1 _Spectrometer_frequency_1H 750 _Mol_system_component_name Arg43Ala-brazzein loop_ _Coupling_constant_code _Atom_one_residue_seq_code _Atom_one_residue_label _Atom_one_name _Atom_two_residue_seq_code _Atom_two_residue_label _Atom_two_name _Coupling_constant_value _Coupling_constant_value_error 3HJNiCj 6 LYS N 49 CYS C 2.89 0.04 3HJNiCj 8 TYR N 47 CYS C 2.80 0.03 3HJNiCj 11 TYR N 46 GLN C 3.00 0.11 3HJNiCj 25 ASP N 21 GLN C 2.88 0.04 3HJNiCj 26 CYS N 22 CYS C 2.79 0.03 3HJNiCj 28 LEU N 24 TYR C 2.88 0.04 3HJNiCj 29 ASP N 25 ASP C 2.81 0.03 3HJNiCj 44 ASN N 41 GLU C 2.82 0.03 3HJNiCj 46 GLN N 38 PHE C 2.86 0.04 3HJNiCj 47 CYS N 9 GLU C 3.00 0.09 3HJNiCj 48 ILE N 36 GLU C 2.89 0.05 3HJNiCj 49 CYS N 6 LYS C 2.98 0.11 3HJNiCj 51 TYR N 4 CYS C 2.80 0.03 stop_ save_